猪源益生芽孢杆菌的分离鉴定与生物学特性分析  

Isolation,Identification and Biological Characterization of Bacillus spp from Swine

作　　者：蒋佳璇 王淑京[2] 任志鸿[2] 杜华茂[1] 

机构地区：[1]西南大学生物技术学院,重庆400715 [2]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,北京102206

出　　处：《动物营养学报》2016年第11期3542-3548,共7页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：传染病预防控制国家重点实验室留学归国人员启动经费(2011SKLl D301)

摘　　要：本研究旨在从健康猪肠道正常菌群中筛选出性能良好的芽孢杆菌,以用于提高饲料利用率、减少猪消化道感染及减少感染后抗生素的使用。本研究精选重庆偏远山区从未添食配合饲料且生产性能优良的经产母猪,从其新鲜粪便中分离耐酸、耐胆盐的芽孢杆菌,进一步筛选能同时产淀粉酶、纤维素酶和蛋白酶的菌株,最后进行生化鉴定和基于16S rDNA序列的分子鉴定。结果表明,试验共筛选出7株能同时分泌淀粉酶、纤维素酶、蛋白酶的芽孢杆菌,它们在pH3.0条件下处理2 h存活率为30%-90%,在0.5%胆盐中处理2 h存活率为40%-100%。经生化鉴定与分子鉴定为1株嗜热脂肪芽孢杆菌,1株地衣芽孢杆菌,5株枯草芽孢杆菌。由结果可知,本研究筛选的7株芽孢杆菌能耐受低pH和高胆盐环境,且具备同时分泌淀粉酶、纤维素酶和蛋白酶3种胞外酶的能力。The aim of this project was to isolate Bacillus spp with good performance from the normal intestinal flora of healthy swine,and to improve the feed utilization rate,reduce the infection of pig digestive tract and reduce the use of antibiotics. The feces specimen were collected from local back-yard cows which never feed with manufactured feeds,and the Bacillus spp with acid and bile salt tolerance was isolated from fresh feces.The isolates were then tested for the production potency of amylase,cellulose and protease on specific agar plates. The strains were identified by biochemical assays and 16 s rDNA sequence alignment analysis. The results showed that 7 Bacillus strains were isolated which could secrete amylase,cellulose and proteinase. The survival ratios of Bacillus strains in pH 3. 0 for 2 hours were 30 % to 90 %,and in 0. 5 % bile salt for 4 hours were 40 % to 100 %. The strains were identified as Bacillus stearothermophiluc（ 1 strain）,Bacillus licheniformis（ 1 strain） and Bacillus subtilis（ 5 strains） by biochemical identification and molecular identification,respectively. In conclusion,it selects 7 strains identified Bacillus spp,which have good tolerance in lowpH and high bile salt conditions,and the strains can secrete amylase,cellulose and proteinase.

关 键 词：益生菌 芽孢杆菌 生化鉴定 16S RDNA 产酶能力 

分 类 号：S811.6[农业科学—畜牧学]